Antioxidant defenses and non-specific immunity at enzymatic and transcriptional levels in response to dietary carbohydrate in a typical carnivorous fish, hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂).

The present study was conducted to explore the influence of dietary carbohydrate on antioxidant capacity and non-specific immunity of hybrid grouper, which would contribute to determine the tolerable dietary carbohydrate content. Seven diets with grade levels of carbohydrate (5.27, 8.95, 11.49, 14.37, 17.78, 20.82 and 23.65%) were fed to triplicate groups of fish for 10 weeks. Results showed that the inclusion of carbohydrate above 11.49% produced significant increased content of hydrogen peroxide (H2O2) in liver and malondialdehyde (MDA) in both serum and liver. The specific activity of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (Gpx) and total antioxidative capacity (T-AOC) were significantly elevated with the increase of dietary carbohydrate from 8.95 to 23.65%, which may be associated with the reduced hepatic soluble protein content. However, opposite variation was observed in the expression of antioxidant related genes (SOD1 and Gpx), which was partly caused by the activation of NF-E2-related factor 2 (Nrf2) and inhibition of Kelch-like-ECH-associated protein 1 (Keap1) at the transcriptional level. The immunoglobulin M (lgM) content and activity of lysozyme and CCP in serum significantly depressed when dietary carbohydrate was above 11.49%. The expression of pro-inflammatory cytokines (TNF-α, IL-1ß and IL-8) was significantly increased with the increase of dietary carbohydrate from 5.27 to 8.95% and thereafter significantly reduced, which was consistent with the changed expression of toll-like receptor 2 (TLR2) and nuclear factor κΒ (NF-κΒ). In above, high dietary carbohydrate significantly impaired the antioxidant capacity and reduced the non-specific immunity of hybrid grouper, and the tolerable dietary carbohydrate content should not exceed 11.49%.

Heterotic patterns of primary and secondary metabolites in the oilseed crop Brassica juncea.

Heterosis refers to the superior performance of F1 hybrids over their respective parental inbred lines. Although the genetic and expression basis of heterosis have been previously investigated, the metabolic basis for this phenomenon is poorly understood. In a preliminary morphological study in Brassica juncea, we observed significant heterosis at the 50% flowering stage, wherein both the growth and reproduction of F1 reciprocal hybrids were greater than that of their parents. To identify the possible metabolic causes or consequences of this heterosis, we carried out targeted LC-MS analysis of 48 primary (amino acids and sugars) and secondary metabolites (phytohormones, glucosinolates, flavonoids, and phenolic esters) in five developmental tissues at 50% flowering in hybrids and inbred parents. Principal component analysis (PCA) of metabolites clearly separated inbred lines from their hybrids, particularly in the bud tissues. In general, secondary metabolites displayed more negative heterosis values in comparison to primary metabolites. The tested primary and secondary metabolites displayed both additive and non-additive modes of inheritance in F1 hybrids, wherein the number of metabolites showing an additive mode of inheritance were higher in buds and siliques (52.77-97.14%) compared to leaf tissues (47.37-80%). Partial least regression (PLS) analysis further showed that primary metabolites, in general, displayed higher association with morphological parameters in F1 hybrids. Overall, our results are consistent with a resource-cost model for heterosis in B. juncea, where metabolite allocation in hybrids appears to favor growth, at the expense of secondary metabolism.

The underlying pathway involved in inter-subspecific hybrid male sterility in rice.

f5 locus in rice (Oryza sativa L.) confers significant effects on hybrid male sterility and segregation distortion. BC14F2 plants with f5-i/i, f5-j/j and f5-i/j genotypes were used to dissect the underlying pathway of f5-caused hybrid male sterility via comparative transcriptome analysis. A total of 350, 421, and 480 differentially expressed genes (DEGs) were identified from f5-i/j vs f5-j/j, f5-j/j vs f5-i/i, and f5-i/j vs f5-i/i, respectively. 145 DEGs were identified simultaneously in f5-i/j vs f5-j/j and f5-i/j vs f5-i/i. Enrichment analysis indicated that stress and cell control related processes were enriched. The expression of ascorbate peroxidase (APX) and most of the heat shock proteins (HSPs) were decreased, which might result in higher sensitivity to various stresses in pollen cells. A model was proposed to summarize the underlying process for f5-caused hybrid male sterility. These results would provide significant clues to further dissecting the molecular mechanism of f5-caused inter-subspecific reproductive isolation.

Productivity and Antioxidant Activity of Wild, Reconstituted, and Hybrid Strains of the Pink Oyster Mushroom, Pleurotus djamor (Agaricomycetes), from Mexico.

The genus Pleurotus is the third most commonly produced edible fungi in the world. In addition, species of genus Pleurotus have functional properties such as anticancer, antiviral, antimicrobial, anti-inflammatory, and antioxidant activities, which are mainly attributed to phenolic compounds. For these reasons, this study evaluated the productivity and antioxidant activity (AA) of 2 wild strains (white and pink), 2 reconstituted strains (called "BB" and "RR"), and 4 hybrid strains (H1, H2, H3, and H4) of P. djamor from monokaryotic components (neohaplonts). The results showed that the white wild-type strain and the reconstituted strains exhibited the best production potential, expressed as biological efficiency and mycelial growth rate. The carpophores of hybrid strains H1 and H3 had the greatest AA, as evaluated with DPPH radical scavenging and reducing power assays, respectively. The H3 strain had the highest total phenol (TP) content. Pearson correlations led us to conclude that the mycelial growth rate has a regular inverse correlation with TP and a regular direct correlation with AA of methanolic extracts from carpophores and myce-lia. This is, to our knowledge, the first report in the literature about the effect of Pleurotus strain hybridization through a chemical de-dikaryotization process on TP content.

Modeling the Evolution of Female Meiotic Drive in Maize.

Autosomal drivers violate Mendel's law of segregation in that they are overrepresented in gametes of heterozygous parents. For drivers to be polymorphic within populations rather than fixing, their transmission advantage must be offset by deleterious effects on other fitness components. In this paper, we develop an analytical model for the evolution of autosomal drivers that is motivated by the neocentromere drive system found in maize. In particular, we model both the transmission advantage and deleterious fitness effects on seed viability, pollen viability, seed to adult survival mediated by maternal genotype, and seed to adult survival mediated by offspring genotype. We derive general, biologically intuitive conditions for the four most likely evolutionary outcomes and discuss the expected evolution of autosomal drivers given these conditions. Finally, we determine the expected equilibrium allele frequencies predicted by the model given recent estimates of fitness components for all relevant genotypes and show that the predicted equilibrium is within the range observed in maize land races for levels of drive at the low end of what has been observed.

Overcoming inter-subspecific hybrid sterility in rice by developing indica-compatible japonica lines.

Rice (Oryza sativa L.) is an important staple crop. The exploitation of the great heterosis that exists in the inter-subspecific crosses between the indica and japonica rice has long been considered as a promising way to increase the yield potential. However, the male and female sterility frequently occurred in the inter-subspecific hybrids hampered the utilization of the heterosis. Here we report that the inter-subspecific hybrid sterility in rice is mainly affected by the genes at Sb, Sc, Sd and Se loci for F1 male sterility and the gene at S5 locus for F1 female sterility. The indica-compatible japonica lines (ICJLs) developed by pyramiding the indica allele (S-i) at Sb, Sc, Sd and Se loci and the neutral allele (S-n) at S5 locus in japonica genetic background through marker-assisted selection are compatible with indica rice in pollen fertility and in spikelet fertility. These results showed a great promise of overcoming the inter-subspecific hybrid sterility and exploiting the heterosis by developing ICJLs.

Cytoplasmic male sterility (CMS) in hybrid breeding in field crops.

KEY MESSAGE: A comprehensive understanding of CMS/Rf system enabled by modern omics tools and technologies considerably improves our ability to harness hybrid technology for enhancing the productivity of field crops. Harnessing hybrid vigor or heterosis is a promising approach to tackle the current challenge of sustaining enhanced yield gains of field crops. In the context, cytoplasmic male sterility (CMS) owing to its heritable nature to manifest non-functional male gametophyte remains a cost-effective system to promote efficient hybrid seed production. The phenomenon of CMS stems from a complex interplay between maternally-inherited (mitochondrion) and bi-parental (nucleus) genomic elements. In recent years, attempts aimed to comprehend the sterility-inducing factors (orfs) and corresponding fertility determinants (Rf) in plants have greatly increased our access to candidate genomic segments and the cloned genes. To this end, novel insights obtained by applying state-of-the-art omics platforms have substantially enriched our understanding of cytoplasmic-nuclear communication. Concomitantly, molecular tools including DNA markers have been implicated in crop hybrid breeding in order to greatly expedite the progress. Here, we review the status of diverse sterility-inducing cytoplasms and associated Rf factors reported across different field crops along with exploring opportunities for integrating modern omics tools with CMS-based hybrid breeding.

[Determination of the composition and the allelic state of disease and pest resistance genes in potato parental lines using DNA markers].

The allelic dosage of disease and pest resistance genes was determined in 11 prospective potato varieties and hybrids by means of detecting the corresponding PCR DNA markers in their progeny from crosses with specially selected testers. It was revealed that most (65%) resistance genes in the analyzed parental lines were present as a single dominant allele (simplex). Nevertheless, we were able to find some multiplex lines valuable for breeding. The Yanka variety and the clone 52-03-16 had one triplex and one duplex of resistance genes, the Lilea and Charaunik varieties and the clone 106-04-17 had two resistance genes in duplex, and the Uladar and Falvarak varieties and the clone 45-04-24 were duplex for some single genes. The highest number of multiplex lines was detected for the genes Ry sto , H1, and Sen1. Only simplex genotypes were revealed for the Gro-1-4 and PLRV1 genes.

Generation of Naïve Bovine Induced Pluripotent Stem Cells Using PiggyBac Transposition of Doxycycline-Inducible Transcription Factors.

Generation of pluripotent stem cells (PSCs) in large domestic animals has achieved only limited success; most of the PSCs obtained to date have been classified as primed PSCs, which possess very little capacity to produce chimeric offspring. By contrast, mouse PSCs have been classified as naïve PSCs that can contribute to most of the tissues of chimeras, including germ cells. Here, we describe the generation of two different types of bovine induced pluripotent stem cells (biPSCs) from amnion cells, achieved through introduction of piggyBac vectors containing doxycycline-inducible transcription factors (Oct3/4, Sox2, Klf4, and c-Myc). One type of biPSCs, cultured in medium supplemented with knockout serum replacement (KSR), FGF2, and bovine leukemia inhibitory factor (bLIF), had a flattened morphology like human PSCs; these were classified as primed-type. The other type biPSCs, cultured in KSR, bLIF, Mek/Erk inhibitor, GSK3 inhibitor and forskolin, had a compact morphology like mouse PSCs; these were classified as naïve-type. Cells could easily be switched between these two types of biPSCs by changing the culture conditions. Both types of biPSCs had strong alkaline phosphatase activity, expressed pluripotent markers (OCT3/4, NANOG, REX1, ESRRß, STELLA, and SOCS3), and formed embryoid bodies that gave rise to differentiated cells from all three embryonic germ layers. However, only naïve-type biPSCs showed the hallmarks of naïve mouse PSCs, such as LIF-dependent proliferation, lack of FGF5 expression, and active XIST expression with two active X chromosomes. Furthermore, naïve-type biPSCs could contribute to the inner cell mass (ICM) of host blastocysts and most tissues within chimeric embryos. This is the first report of generation of biPSCs with several characteristics similar to those of naïve mouse PSCs and a demonstrated potential to contribute to chimeras.

Salt stress response of membrane proteome of sugar beet monosomic addition line M14.

Understanding how plants respond to and tolerate salt stress is important for engineering and breeding effort to boost plant productivity and bioenergy in an ever challenging environment. Sugar beet M14 line is a unique germplasm that contains genetic materials from Beta vulgaris L. and Beta corolliflora Zoss, and it exhibits tolerance to salt stress. Here we report the changes in membrane proteome of the M14 plants in response to salt stress (0, 200, 400mM NaCl) using an iTRAQ two-dimensional LC-MS/MS technology for quantitative proteomic analysis. In total, 274 proteins, mostly membrane proteins, were identified, and 50 proteins exhibited differential protein level changes, with 40 proteins increased and 10 decreased. The proteins were mainly involved in transport, metabolism, protein synthesis, photosynthesis, protein folding and degradation, signal transduction, stress and defense, energy, and cell structure. These results have revealed interesting mechanisms underlying the M14 response and tolerance to salt stress. BIOLOGICAL SIGNIFICANCE: Sugar beet monosomic addition line M14 is a special variety with salt stress tolerance. Analysis of the M14 membrane proteome under salt stress may provide useful information regarding specific adaptive mechanisms underlying salt stress tolerance. Membrane proteins are known to play critical roles in salt stress signaling and adaptation. The purpose of this study was to identify significantly changed membrane proteins and determine their possible relevance to salt tolerance. The proteomic analysis of the M14 line revealed important molecular mechanisms that can be potentially applied to improving crop salt tolerance. This article is part of a Special Issue entitled: Proteomics in India.

Genotypic variability for antioxidant and quality parameters among tomato cultivars, hybrids, cherry tomatoes and wild species.

BACKGROUND: Wide germplasm diversity and transferability of antioxidant parameters is the primary requirement for the development of high-antioxidant tomato cultivars. The present study was conducted to screen tomato genotypes including hybrids, varieties, cherry tomatoes, wild species, elite germplasm lines, interspecific hybrids and backcross populations for antioxidant activity and other quality parameters to select high-antioxidant lines with good total soluble solids (TSS) for further usage in crop improvement programmes. RESULTS: Wild species and interspecific hybrids between LA-1777 (Solanum habrochaites) and an elite genotype 15SBSB recorded very high antioxidant capacity (FRAP), DPPH radical-scavenging ability, and high phenols and flavonoids. Interspecific hybrids also recorded very high total soluble solids (TSS). Significantly higher total carotenoids, lycopene and vitamin C were observed in IIHR-249-1 with moderately higher TSS. Cherry tomato lines IIHR-2866, 2865 and 2864 recorded four to five times more ß-carotene than commercial hybrids/varieties. CONCLUSION: Tomato line IIHR-249-1 can be used for improving antioxidant capacity, total carotenoids and lycopene in tomato breeding programmes. Cherry tomato lines IIHR-2866, 2865 and 2864 can be used for improving ß-carotene content. LA-1777 and interspecific hybrids could be used for developing tomato lines rich in antioxidants as well as TSS.

[Effect of Triton X-100 on genetic segregation and associated monocotyledonous and dicotyledonous traits in sugarbeet (Beta vulgaris L.)].

The effect of Triton X-100 (TX-100) on the ratio of phenotypic classes and the expression of morphological traits in the progeny of sugar beet hybrids (N12 and N2) was investigated. It was shown that the TX-100 exposition on the unopened flower buds of sugar beets has different effects on hybrid progenies. In agamospermic progeny of hybrid plant No 12km-4, a significant decrease in the heteroallelic (heterozygous) phenotypic classes of alcohol dehydrogenase (ADH1) fraction was determined in the nonagamospermic progeny of hybrid plant No 2km-2 appearance of sugar beet seedlings with one cotyledon leaf was detected. The obtained results indicate the high efficiency of the epimutagenic effect of TX-100 on the early stages of plant ontogenesis.

Preservation of Solanum pimpinellifolium genomic fragments in recombinant genotypes improved the fruit quality of tomato.

Five recombinant inbred lines obtained from the F2 generation of an interspecific cross between cultivar, Caimanta (Cai, Solanum lycopersicum) and wild accession, LA722 (P, S. pimpinellifolium) were crossed to obtain the second cycle hybrids (SCH). Eleven fruit quality traits were assessed in evaluating phenotypic variability among genotypes P, Cai, F1 (Cai x P), five RILs, and 10 SCH. One of the five recombinant inbred lines and three SCH had higher values than P, as the best genotype for shelf life. Sequence-related amplified polymorphism was used as the molecular method for detecting polymorphism among these 18 genotypes. The percentage of polymorphism in RILs and SCH was 61% and 66% respectively. Moreover, some bands detected in P were present in SCH. Several multivariate analyses were performed to find agreement between the phenotypic variability observed for fruit quality traits and the polymorphism obtained from sequence-related amplified polymorphism markers. A general Procrustes analysis estimated that there was a consensus proportion of 75% between phenotypic and molecular data. There was considerable preservation of some bands from the wild genotype, which could increase the variability in fruit quality traits in populations where the genetic diversity is limited.

Preliminary genetic linkage maps of Chinese herb Dendrobium nobile and D. moniliforme.

Dendrobium is an endangered genus in the orchid family with medicinal and horticultural value. Two preliminary genetic linkage maps were constructed using 90 F1 progeny individuals derived from an interspecific cross between D. nobile and D. moniliforme (both, 2n = 38), using random amplified polymorphic DNA (RAPD) and intersimple sequence repeat (ISSR). A total of 286 RAPD loci and 68 ISSR loci were identified and used for genetic linkage analysis. Maps were constructed by double pseudo-testcross mapping strategy using the software Mapmaker/EXP ver. 3.0, and Kosambi map distances were constructed using a LOD score greater than or equal to 4 and a recombination threshold of 0.4. The resulting frame map of D. nobile was 1474 cM in total length with 116 loci distributed in 15 linkage groups; and the D. moniliforme linkage map had 117 loci placed in 16 linkage groups spanning 1326.5 cM. Both maps showed 76.91% and 73.59% genome coverage for D. nobile and D. moniliforme, respectively. These primary maps provide an important basis for genetic studies and further medicinal and horticultural traits mapping and marker-assisted selection in Dendrobium breeding programmes.

Assessing the bioconfinement potential of a Nicotiana hybrid platform for use in plant molecular farming applications.

BACKGROUND: The introduction of pharmaceutical traits in tobacco for commercial production could benefit from the utilization of a transgene bioconfinement system. It has been observed that interspecific F1Nicotiana hybrids (Nicotiana tabacum × Nicotiana glauca) are sterile and thus proposed that hybrids could be suitable bioconfined hosts for biomanufacturing. We genetically tagged hybrids with green fluorescent protein (GFP), which was used as a visual marker to enable gene flow tracking and quantification for field and greenhouse studies. GFP was used as a useful proxy for pharmaceutical transgenes. RESULTS: Analysis of DNA content revealed significant genomic downsizing of the hybrid relative to that of N. tabacum. Hybrid pollen was capable of germination in vitro, albeit with a very low frequency and with significant differences between plants. In two field experiments, one each in Tennessee and Kentucky, we detected outcrossing at only one location (Tennessee) at 1.4%. Additionally, from 50 hybrid plants at each field site, formation of 84 and 16 seed was observed, respectively. Similar conclusions about hybrid fertility were drawn from greenhouse crosses. In terms of above-ground biomass, the hybrid yield was not significantly different than that of N. tabacum in the field. CONCLUSION: N. tabacum × N. glauca hybrids show potential to contribute to a bioconfinement- and biomanufacturing host system. Hybrids exhibit extremely low fertility with no difference of green biomass yields relative to N. tabacum. In addition, hybrids are morphologically distinguishable from tobacco allowing for identity preservation. This hybrid system for biomanufacturing would optimally be used where N. glauca is not present and in physical isolation of N. tabacum production to provide total bioconfinement.

Melatonin improves the reprogramming efficiency of murine-induced pluripotent stem cells using a secondary inducible system.

This study focused on the effect of melatonin on reprogramming with specific regard to the generation of induced pluripotent stem cells (iPSCs). Here, a secondary inducible system, which is more accurate and suitable for studying the involvement of chemicals in reprogramming efficiency, was used to evaluate the effect of melatonin on mouse iPSC generation. Secondary fibroblasts collected from all-iPSC mice through tetraploid complementation were cultured in induction medium supplemented with melatonin at different concentrations (0, 10(-6), 10(-7), 10(-8), 10(-9), or 10(-10 )m) or with vitamin C (50 µg/mL) as a positive control. Compared with untreated group (0.22 ± 0.04% efficiency), 10(-8) (0.81 ± 0.04%), and 10(-9 )m (0.83 ± 0.08%) melatonin supplementation significantly improved reprogramming efficiency (P < 0.05). Moreover, we verified that the iPSCs induced by melatonin treatment (MiPSCs) had the same characteristics as typical embryonic stem cells (ESCs), including expression of the pluripotency markers Oct4, Sox2, and Nanog, the ability to form teratomas and all three germ layers of the embryo, as well as produce chimeric mice with contribution to the germ line. Interestingly, only the melatonin receptor MT2 was detected in secondary fibroblasts, while MiPSCs and ESCs expressed MT1 and MT2 receptors. Furthermore, during the early stage of reprogramming, expression of the apoptosis-related genes p53 and p21 was lower in the group treated with 10(-9) m melatonin compared with the untreated controls. In conclusion, melatonin supplementation enhances the efficiency of murine iPSC generation. These beneficial effects may be associated with inhibition of the p53-mediated apoptotic pathway.

Wnt signal specifies the intrathalamic limit and its organizer properties by regulating Shh induction in the alar plate.

The structural complexity of the brain depends on precise molecular and cellular regulatory mechanisms orchestrated by regional morphogenetic organizers. The thalamic organizer is the zona limitans intrathalamica (ZLI), a transverse linear neuroepithelial domain in the alar plate of the diencephalon. Because of its production of Sonic hedgehog, ZLI acts as a morphogenetic signaling center. Shh is expressed early on in the prosencephalic basal plate and is then gradually activated dorsally within the ZLI. The anteroposterior positioning and the mechanism inducing Shh expression in ZLI cells are still partly unknown, being a subject of controversial interpretations. For instance, separate experimental results have suggested that juxtaposition of prechordal (rostral) and epichordal (caudal) neuroepithelium, anteroposterior encroachment of alar lunatic fringe (L-fng) expression, and/or basal Shh signaling is required for ZLI specification. Here we investigated a key role of Wnt signaling in the molecular regulation of ZLI positioning and Shh expression, using experimental embryology in ovo in the chick. Early Wnt expression in the ZLI regulates Gli3 and L-fng to generate a permissive territory in which Shh is progressively induced by planar signals of the basal plate.

[Study of the effects of gamma-irradiation of common wheat F1 seeds using gliadins as genetic markers].

Effects of irradiation of dry F1 seeds with gamma rays in the dose of 200 Gy were studied. Hybrids between near-isogenic lines on the basis of the variety Bezostaya 1 served as the material of investigation. Irradiation markedly reduced productivity traits of F1 plants and did not affect the survival of F1 plants under the given growth conditions. A significant relative increase in the frequency of pollen grains with the 1BL/1RS translocation that formed F2 seeds in comparison with the control was one of the effects of irradiation of F1 seeds. Irradiation with gamma-rays induced mutations at gliadin loci with the frequency of 7,4 % (at 0,5 % in the control).

Cytogenetic studies on meiotic chromosome behaviors in sterile Oriental x Trumpet lily.

In order to determine the reasons for pollen sterility in lily hybrids, four diploid sterile Oriental x Trumpet (OT) lily cultivars ('Nymph', 'Gluhwein', 'Yelloween', and 'Shocking') were used to investigate the meiotic chromosome behaviors in pollen mother cells (PMCs), using genomic in situ hybridization and conventional cytological methods. At metaphase I, chromosome associations were quite variable, not only among different genotypes but also in different PMCs of the same genotype. In addition to bivalents, a certain amount of univalent, trivalents, and quadrivalents were observed in all of the investigated genotypes. In addition, ring octavalents and ring hexavalents were observed in 'Nymph'. Even dodecavalents were observed in 'Nymph'. These abnormal chromosome associations at metaphase I implied the occurrence of chromosome interchanges (translocation) in these intersectional hybrids. At anaphase-telophase, a large number of laggard chromosomes and different kinds of chromosome bridge configurations were observed. At the tetrad stage, micronuclei and polyads were also found in many PMCs. All of these abnormal chromosome behaviors in PMCs were responsible for the pollen sterility in lily hybrids.

Selection and authentication of botanical materials for the development of analytical methods.

Herbal products, for example botanical dietary supplements, are widely used. Analytical methods are needed to ensure that botanical ingredients used in commercial products are correctly identified and that research materials are of adequate quality and are sufficiently characterized to enable research to be interpreted and replicated. Adulteration of botanical material in commerce is common for some species. The development of analytical methods for specific botanicals, and accurate reporting of research results, depend critically on correct identification of test materials. Conscious efforts must therefore be made to ensure that the botanical identity of test materials is rigorously confirmed and documented through preservation of vouchers, and that their geographic origin and handling are appropriate. Use of material with an associated herbarium voucher that can be botanically identified is always ideal. Indirect methods of authenticating bulk material in commerce, for example use of organoleptic, anatomical, chemical, or molecular characteristics, are not always acceptable for the chemist's purposes. Familiarity with botanical and pharmacognostic literature is necessary to determine what potential adulterants exist and how they may be distinguished.